title: Blue light regulated two-component systems: enzymatic and functional analyses of light-oxygen-voltage (LOV)-histidine kinases and downstream response regulators.
abstract: Light is an essential environmental cue for diverse organisms. Many prokaryotic blue light photoreceptors use light, oxygen, voltage (LOV) sensory domains to control the activities of diverse output domains, including histidine kinases (HK). Upon activation, these proteins autophosphorylate a histidine residue before subsequently transferring the phosphate to an aspartate residue in the receiver domain of a cognate response regulator (RR). Such phosphorylation activates the output domain of the RR, leading to changes in gene expression, protein-protein interactions, or enzymatic activities. Here, we focus on one such light sensing LOV-HK from the marine bacterium Erythrobacter litoralis HTCC2594 (EL368), seeking to understand how kinase activity and subsequent downstream effects are regulated by light. We found that photoactivation of EL368 led to a significant enhancement in the incorporation of phosphate within the HK domain. Further enzymatic studies showed that the LOV domain affected both the LOV-HK turnover rate (kcat) and Km in a light-dependent manner. Using in vitro phosphotransfer profiling, we identified two target RRs for EL368 and two additional LOV-HKs (EL346 and EL362) encoded within the host genome. The two RRs include a PhyR-type transcriptional regulator (EL_PhyR) and a receiver-only protein (EL_LovR), reminiscent of stress-triggered systems in other bacteria. Taken together, our data provide a biochemical foundation for this light-regulated signaling module of sensors, effectors, and regulators that control bacterial responses to environmental conditions.
authors.affiliation: Departments of Biophysics and Biochemistry, University of Texas Southwestern Medical Center , Dallas, Texas 75390, United States.
authors.forename: Fernando
authors.initials: F
authors.lastname: Correa
authors.forename: Wen-Huang
authors.initials: WH
authors.lastname: Ko
authors.forename: Victor
authors.initials: V
authors.lastname: Ocasio
authors.forename: Roberto A
authors.initials: RA
authors.lastname: Bogomolni
authors.forename: Kevin H
authors.initials: KH
authors.lastname: Gardner
mesh_terms: D002850:Chromatography, Gel; D003001:Cloning, Molecular; D019281:Dimerization; D000071677:Histidine Kinase; D007700:Kinetics; D008027:Light; D008958:Models, Molecular; D010100:Oxygen / Q000737:chemistry*; D010766:Phosphorylation; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics; D012542:Scattering, Radiation; D015398:Signal Transduction
chemical_list: D011494:Protein Kinases; D000071677:Histidine Kinase; D010100:Oxygen